动物源性肠出血性大肠杆菌O157∶H7及其3个毒力基因
的多重PCR快速检测研究
陈雅君;王亚宾;张莉娟;张龙现;陈丽颖;刘中原;申果;胡慧
【期刊名称】《中国人兽共患病学报》 【年(卷),期】2013(029)007
【摘要】目的 建立快速、特异的分离鉴定大肠杆菌O157∶H7及其3个主要毒力基因(hylA、eaeA和stx2基因)的多重PCR方法.方法 根据GenBank公布的大肠杆菌O157∶H7菌体抗原rfbE基因、鞭毛抗原fliC基因、溶血素(hlyA)基因、紧密黏附素(eaeA)基因和志贺样毒素2(stx2)基因为靶基因,设计5对特异性引物,在同一扩增体系中进行PCR,优化反应体系,测定特异性和灵敏度,并进行了临床样品的检测.结果 该方法扩增目的基因片段分别为327 bp、247 bp、494 bp、384bp和779 bp,特异性和灵敏度均高,细菌纯培养物的检测灵敏度为104 cfu/mL.结论 初步建立了快速、灵敏、特异的检测肠出血性大肠杆菌O157∶H7及其3个毒力基因的多重PCR方法,可用于临床动物携带大肠杆菌O157∶H7的分子流行病学调查及食品微生物检测.%A multiplex PCR method was developed for rapid and specific detection of Escherichia coli O157∶H7 and three of its virulence genes (hylA,eaeA and stx2 genes).Five sets of primers were designed according to the sequences of rfbE gene,fliC gene,hemolysin gene (hlyA),inthnin gene (eaeA),and shiga-like toxin gene 2(stx2) of E.coli O157.They were selected and added into one amplification system to perform PCR.The system was optimized,and the specificity and sensitivity of this system were
动物源性肠出血性大肠杆菌O157∶H7及其3个毒力基因的多重PCR快速检测研究
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